Details
| Original language | English |
|---|---|
| Pages (from-to) | 3671-3680 |
| Number of pages | 10 |
| Journal | ACS Nano |
| Volume | 11 |
| Issue number | 4 |
| Publication status | Published - 14 Mar 2017 |
Abstract
Efficiently delivering functional cargo to millions of cells on the time scale of minutes will revolutionize gene therapy, drug discovery, and high-throughput screening. Recent studies of intracellular delivery with thermoplasmonic structured surfaces show promising results but in most cases require time- or cost-intensive fabrication or lead to unreproducible surfaces. We designed and fabricated large-area (14 × 14 mm), photolithography-based, template-stripped plasmonic substrates that are nanosecond laser-activated to form transient pores in cells for cargo entry. We optimized fabrication to produce plasmonic structures that are ultrasmooth and precisely patterned over large areas. We used flow cytometry to characterize the delivery efficiency of cargos ranging in size from 0.6 to 2000 kDa to cells (up to 95% for the smallest molecule) and viability of cells (up to 98%). This technique offers a throughput of 50000 cells/min, which can be scaled up as necessary. This technique is also cost-effective as each large-area photolithography substrate can be used to deliver cargo to millions of cells, and switching to a nanosecond laser makes the setup cheaper and easier to use. The approach we present offers additional desirable features: spatial selectivity, reproducibility, minimal residual fragments, and cost-effective fabrication. This research supports the development of safer genetic and viral disease therapies as well as research tools for fundamental biological research that rely on effectively delivering molecules to millions of living cells.
Keywords
- flow cytometry, plasmonic intracellular delivery, pulsed lasers, template-stripping, thermoplasmonic substrates
ASJC Scopus subject areas
- Materials Science(all)
- General Materials Science
- Engineering(all)
- General Engineering
- Physics and Astronomy(all)
- General Physics and Astronomy
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In: ACS Nano, Vol. 11, No. 4, 14.03.2017, p. 3671-3680.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Intracellular Delivery Using Nanosecond-Laser Excitation of Large-Area Plasmonic Substrates
AU - Saklayen, Nabiha
AU - Huber, Marinus
AU - Madrid, Marinna
AU - Nuzzo, Valeria
AU - Vulis, Daryl I.
AU - Shen, Weilu
AU - Nelson, Jeffery
AU - McClelland, Arthur A.
AU - Heisterkamp, Alexander
AU - Mazur, Eric
N1 - Funding information: The research described in this paper was supported by the National Science Foundation under contracts PHY-1219334 and PHY-1205465. N.S. was funded by the Howard Hughes Medical Institute's International Fellowship. M.M. was funded by the Graduate Prize Fellowship at Harvard University. A.H. received funding from the German Research Foundation through the Cluster of Excellence REBIRTH (DFG EXC62/3).
PY - 2017/3/14
Y1 - 2017/3/14
N2 - Efficiently delivering functional cargo to millions of cells on the time scale of minutes will revolutionize gene therapy, drug discovery, and high-throughput screening. Recent studies of intracellular delivery with thermoplasmonic structured surfaces show promising results but in most cases require time- or cost-intensive fabrication or lead to unreproducible surfaces. We designed and fabricated large-area (14 × 14 mm), photolithography-based, template-stripped plasmonic substrates that are nanosecond laser-activated to form transient pores in cells for cargo entry. We optimized fabrication to produce plasmonic structures that are ultrasmooth and precisely patterned over large areas. We used flow cytometry to characterize the delivery efficiency of cargos ranging in size from 0.6 to 2000 kDa to cells (up to 95% for the smallest molecule) and viability of cells (up to 98%). This technique offers a throughput of 50000 cells/min, which can be scaled up as necessary. This technique is also cost-effective as each large-area photolithography substrate can be used to deliver cargo to millions of cells, and switching to a nanosecond laser makes the setup cheaper and easier to use. The approach we present offers additional desirable features: spatial selectivity, reproducibility, minimal residual fragments, and cost-effective fabrication. This research supports the development of safer genetic and viral disease therapies as well as research tools for fundamental biological research that rely on effectively delivering molecules to millions of living cells.
AB - Efficiently delivering functional cargo to millions of cells on the time scale of minutes will revolutionize gene therapy, drug discovery, and high-throughput screening. Recent studies of intracellular delivery with thermoplasmonic structured surfaces show promising results but in most cases require time- or cost-intensive fabrication or lead to unreproducible surfaces. We designed and fabricated large-area (14 × 14 mm), photolithography-based, template-stripped plasmonic substrates that are nanosecond laser-activated to form transient pores in cells for cargo entry. We optimized fabrication to produce plasmonic structures that are ultrasmooth and precisely patterned over large areas. We used flow cytometry to characterize the delivery efficiency of cargos ranging in size from 0.6 to 2000 kDa to cells (up to 95% for the smallest molecule) and viability of cells (up to 98%). This technique offers a throughput of 50000 cells/min, which can be scaled up as necessary. This technique is also cost-effective as each large-area photolithography substrate can be used to deliver cargo to millions of cells, and switching to a nanosecond laser makes the setup cheaper and easier to use. The approach we present offers additional desirable features: spatial selectivity, reproducibility, minimal residual fragments, and cost-effective fabrication. This research supports the development of safer genetic and viral disease therapies as well as research tools for fundamental biological research that rely on effectively delivering molecules to millions of living cells.
KW - flow cytometry
KW - plasmonic intracellular delivery
KW - pulsed lasers
KW - template-stripping
KW - thermoplasmonic substrates
UR - http://www.scopus.com/inward/record.url?scp=85018652151&partnerID=8YFLogxK
U2 - 10.1021/acsnano.6b08162
DO - 10.1021/acsnano.6b08162
M3 - Article
C2 - 28291329
AN - SCOPUS:85018652151
VL - 11
SP - 3671
EP - 3680
JO - ACS Nano
JF - ACS Nano
SN - 1936-0851
IS - 4
ER -